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miR-146a promotes M2 macrophage polarization and accelerates diabetic wound healing by inhibiting the TLR4/NF-κB axis
in Journal of Molecular Endocrinology*(X Peng and F He contributed equally to this work)
We tried to unveil the clinical significance of miR-146a as a biomarker in M2 macrophage polarization in diabetic wound healing. Initially, we found reduced miR-146a in macrophages of diabetic patients. Next, dual-luciferase assay verified that toll-like receptor 4 (TLR4) was a target gene of miR-146 and was negatively regulated by miR-146. Moreover, after ectopic expression and depletion experiments of miR-146 and/or TLR4, lipopolysaccharide-induced inflammatory response of macrophages was detected. The results revealed that overexpression of miR-146a promoted the M2 macrophage polarization by suppressing the TLR4/nuclear factor-kappaB (NF-κB) axis, so as to enhance wound healing in diabetic ulcers. Further, mouse models with diabetic ulcers were established to investigate the effects of miR-146a on diabetic wound healing in vivo, which revealed that miR-146a promoted wound healing in diabetic ulcers by inhibiting the TLR4/NF-κB axis. In conclusion, we demonstrate that miR-146a can induce M2 macrophage polarization to enhance wound healing in diabetic ulcers by inhibiting the TLR4/NF-κB axis.
Supplementary Materials
- Supplementary Figure 1 A, Flow cytometric analysis of M1 cell surface marker proteins CD86 and CD40. B, Flow cytometric analysis of M2 cell surface marker proteins CD163 and CD206. C, Flow cytometric analysis of CD163 and CD206 proteins in diabetic mice.
- Supplementary Figure 2 A, Representative image of protein band of IkB phosphorylation and expression in macrophages detected by Western blot analysis. B, Representative image of protein band of the phosphorylation level of p65 in macrophages detected by Western blot analysis. C, Representative image of protein band of TLR4 expression in macrophages treated with siRNA-TLR4 determined using Western blot analysis. D, Representative image of protein band of phosphorylation level of p65 in macrophages
- Supplementary Table 1 RT-qPCR primer sequences (mice)
- Supplementary Table 2 RT-qPCR primer sequences (human)
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